Predictive Modeling for Growth of Non‐ and Cold‐adapted Listeria monocytogenes on Fresh‐cut Cantaloupe at Different Storage Temperatures

The aim of this study was to determine the growth kinetics of Listeria monocytogenes, with and without cold‐adaption, on fresh‐cut cantaloupe under different storage temperatures. Fresh‐cut samples, spot inoculated with a 4‐strain cocktail of L. monocytogenes (～3.2 log CFU/g), were exposed to constant storage temperatures held at 10, 15, 20, 25, or 30 °C. All growth curves of L. monocytogenes were fitted to the Baranyi, modified Gompertz, and Huang models. Regardless of conditions under which cells grew, the time needed to reach 5 log CFU/g decreased with the elevated storage temperature. Experimental results showed that there were no significant differences (P > 0.05) in the maximum growth rate k (log CFU/g h^?1) and lag phase duration λ (h) between the cultures of L. monocytogenes with or without previous cold‐adaption treatments. No distinct difference was observed in the growth pattern among 3 primary models at various storage temperatures. The growth curves of secondary modeling were fitted on an Arrhenius‐type model for describing the relationship between k and temperature of the L. monocytogenes on fresh‐cut cantaloupe from 10 to 30 °C. The root mean square error values of secondary models for non‐ and cold‐adapted cells were 0.018, 0.021, and 0.024, and 0.039, 0.026, and 0.017 at the modified Gompertz, Baranyi, and Huang model, respectively, indicating that these 3 models presented the good statistical fit. This study may provide valuable information to predict the growth of L. monocytogenes on fresh‐cut cantaloupes at different storage conditions.     

Growth Potential of Listeria Monocytogenes and Staphylococcus Aureus on Fresh‐Cut Tropical Fruits

The objective of this study was to evaluate the fate of Staphylococcus aureus, Listeria monocytogenes, and natural microbiota on fresh‐cut tropical fruits (pitaya, mango, papaya and pineapple) with commercial PVC film at different storage temperature (5, 13, and 25 °C). The results showed that S. aureus, L. monocytogenes, and natural microbiota increased significantly on fresh‐cut tropical fruits at 25 °C. Both pathogen and natural microbiota were able to grow on fresh‐cut tropical fruits at 13 °C. The maximum population of L. monocytogenes was higher than that of S. aureus on fresh‐cut tropical fruits. L. monocytogenes and S. aureus could survive without growth on fresh‐cut pitaya, mango, and papaya at 5 °C. The population of L. monocytogenes declined significantly on fresh‐cut pineapple at all temperature, indicating composition of fresh‐cut pineapple could inhibit growth of L. monocytogenes. However, S. aureus was still able to grow on fresh‐cut pineapple at storage temperature. Thus, this study suggests that 4 kinds of fresh‐cut tropical fruits (pitaya, mango, papaya, and pineapple) should be stored at low temperature to extend shelf life as well as to ensure the safety of fresh‐cut fruits.     

A Rapid and Simple DNA Extraction Procedure to Detect Salmonella spp. and Listeria monocytogenes from Fresh Produce Using Real-time PCR

DNA isolation procedures significantly influence the outcome of PCR-based detection of human pathogens. Unlike clinical samples, DNA isolation from food samples, particularly from fresh and fresh-cut produce has remained a formidable task and has hampered the sensitivity and accuracy of molecular methods. We utilized a commercially available filter-based DNA isolation method (FTA) in conjunction with real-time PCR-based detection of Salmonella spp. and Listeria monocytogenes. The protocol uses filter paper discs impregnated with a patented chemical formulation that lyses cells, immobilizes DNA, and protects it from degradation. Use of the FTA method in conjunction with real-time PCR for the detection of Salmonella spp. and L. monocytogenes was compared with two commercially available DNA isolation procedures that are commonly used for high throughput real-time PCR pathogen detection systems. Both pathogens were successfully detected from artificially inoculated fresh and fresh-cut produce such as alfalfa sprouts, cilantro, green onion, broccoli, prepacked mixed salad, and spinach at low cell numbers (four to seven colony forming units per 25 g initial inoculum level before enrichment). The FTA protocol had distinct advantages of simplicity, biosafety, and compatibility for high throughput screening. This DNA preparation protocol was rapid, sensitive, required minimal handling, and reduced interference from produce-associated inhibitors of real-time PCR. Mention of brand names does not constitute an endorsement by the U.S. Department of Agriculture above others of a similar nature not mentioned.     

Multiplex PCR identification of Listeria monocytogenes isolates from milk and milk‐processing environments

A multiplex PCR procedure based on genes iap (coding for the invasion‐associated 60 kDa protein or p60) and hly (coding for listeriolysin O) of Listeria monocytogenes was to used to investigate the status of its contamination along the major milk‐processing environments. Duplex PCR amplified fragments of the iap gene at about 1.45 kb from all strains of major Listeria spp. tested and a 420 bp fragment of hly from L. monocytogenes reference strains. With triplex PCR, all L. monocytogenes strains exhibited a 420 bp fragment of hly as well as a 700 bp fragment of iap instead of its 1.45kb PCR product. The tentative L. monocytogenes isolates (n = 27) out of 566 samples of milk and milk‐processing environments in the PALCAM agar from cultures of buffered listeria enrichment broth were further subjected to both API and triplex PCR identification. Both methods identified the same 20 isolates as L. monocytogenes. The triplex PCR procedure detected as low as 3.2 × 10¹ cfu mL^?1 of listerial cells even in the presence of L. innocua (10⁸ cfu mL^?1). It could detect 1.4 × 10² listerial cells mL^?1 directly from milk artificially contaminated with the bacteria. Lower levels of L. monocytogenes cells in milk (1.45 × 10¹ to 1.45 × 10⁰ cfu mL^?1) could be detectable if the inoculated milk samples were cultured for 3–6 h. The bacterium was found not only in raw milk, sewage water and vessel surfaces but also in pasteurized milk. Copyright © 2005 Society of Chemical Industry     

Combined Effect of Thermosonication and Slightly Acidic Electrolyzed Water to Reduce Foodborne Pathogens and Spoilage Microorganisms on Fresh‐cut Kale

This study evaluated the efficacy of individual treatments (thermosonication [TS+DW] and slightly acidic electrolyzed water [SAcEW]) and their combination on reducing Escherichia coli O157:H7, Listeria monocytogenes, and spoilage microorganisms (total bacterial counts [TBC], Enterobacteriaceae, Pseudomonas spp., and yeast and mold counts [YMC]) on fresh‐cut kale. For comparison, the antimicrobial efficacies of sodium chlorite (SC; 100 mg/L) and sodium hypochlorite (SH; 100 mg/L) were also evaluated. Each 10 g sample of kale leaves was inoculated to contain approximately 6 log CFU/g of E. coli O157:H7 or L. monocytogenes. Each inoculated or uninoculated samples was then dip treated with deionized water (DW; control), TS+DW, and SAcEW at various treatment conditions (temperature, physicochemical properties, and time) to assess the efficacy of each individual treatment. The efficacy of TS+DW or SAcEW was enhanced at 40 °C for 3 min, with an acoustic energy density of 400 W/L for TS+DW and available chlorine concentration of 5 mg/L for SAcEW. At 40 °C for 3 min, combined treatment of thermosonication 400 W/L and SAcEW 5 mg/L (TS+SAcEW) was more effective in reducing microorganisms compared to the individual treatments (SAcEW, SC, SH, and TS+DW) and combined treatments (TS+SC and TS+SH), which significantly (P < 0.05) reduced E. coli O157:H7, L. monocytogenes, TBC, Enterobacteriaceae, Pseudomonas spp., and YMC by 3.32, 3.11, 3.97, 3.66, 3.62, and >3.24 log CFU/g, respectively. The results suggest that the combined treatment of TS+SAcEW has the potential as a decontamination process in fresh‐cut industry.     

Effect of pomegranate (Punica granatum) pomace extract as a washing agent on the inactivation of Listeria monocytogenes inoculated on fresh produce

Pomegranate pomace extract (PPE) was used as a substitute to chlorinated water for washing of fresh produce. PPE contained punicalagins and ellagic acids, and it had a good inhibitory effect against Listeria monocytogenes based on the results of disc diffusion test and time‐kill assay. Red chard, beet, chicon and red mustard leaves were treated with 7 mg mL^?1 PPE as a washing solution to inactivate L. monocytogenes inoculated on fresh produce and compared with 0.05 mg mL^?1 NaOCl treatment. After the treatment, L. monocytogenes population on the four leafy vegetables was reduced by 1.96–2.97 log CFU g^?1 compared to that on nontreated samples. The reductions in L. monocytogenes after PPE treatment were higher than those observed after NaOCl treatment. Further, the PPE treatment did not affect the surface colour of all fresh produce applied in this study. Thus, PPE can be used as an effective antimicrobial washing agent of fresh produce.     

A Comparative Study of Natural Antimicrobial Delivery Systems for Microbial Safety and Quality of Fresh‐Cut Lettuce

Nanoencapsulation can provide a means to effectively deliver antimicrobial compounds and enhance the safety of fresh produce. However, to date there are no studies which directly compares how different nanoencapsulation systems affect fresh produce safety and quality. This study compared the effects on quality and safety of fresh‐cut lettuce treated with free and nanoencapsulated natural antimicrobial, cinnamon bark extract (CBE). A challenge study compared antimicrobial efficacy of 3 different nanoencapsulated CBE systems. The most effective antimicrobial treatment against Listeria monocytogenes was chitosan‐co‐poly‐N‐isopropylacrylamide (chitosan‐PNIPAAM) encapsulated CBE, with a reduction on bacterial load up to 2 log₁₀ CFU/g (P < 0.05) compared to the other encapsulation systems when fresh‐cut lettuce was stored at 5 °C and 10 °C for 15 d. Subsequently, chitosan‐PNIPAAM‐CBE nanoparticles (20, 40, and 80 mg/mL) were compared to a control and free CBE (400, 800, and 1600 μg/mL) for its effects on fresh‐cut lettuce quality over 15 d at 5 °C. By the 10th day, the most effective antimicrobial concentration was 80 mg/mL for chitosan‐PNIPAAM‐CBE, up to 2 log₁₀ CFU/g reduction (P < 0.05), compared with the other treatments. There was no significant difference between control and treated samples up to day 10 for the quality attributes evaluated. Chitosan‐PNIPAAM‐CBE nanoparticles effectively inhibited spoilage microorganisms’ growth and extended fresh‐cut lettuce shelf‐life. Overall, nanoencapsulation provided a method to effectively deliver essential oil and enhanced produce safety, while creating little to no detrimental quality changes on the fresh‐cut lettuce.     

Survival and Metabolic Activity of Listeria monocytogenes on Ready‐to‐Eat Roast Beef Stored at 4 °C

Three brands of commercial roast beef were purchased and artificially inoculated with a 5‐strain Listeria monocytogenes cocktail at 2 inoculation levels (approximately 3 and 6 Log CFU/g). Although all 3 brands contained sodium diacetate and sodium lactate, inoculated Listeria cocktail survived for 16 d in all 3 brands; significant increases in L. monocytogenes numbers were seen on inoculated Brand B roast beef on days 12 and 16. Numbers of L. monocytogenes increased to 4.14 Log CFU/g for the 3 Log CFU/g inoculation level and increased to 7.99 Log CFU/g for the 6 Log CFU/g inoculation level by day 16, with the pH values being 5.4 and 5.8 respectively. To measure the cell viability in potential biofilms formed, an Alamar blue assay was conducted. Brand B meat homogenate had the highest metabolic activities (P < 0.05). By comparing its metabolic activities to Brands A and C and the inoculated autoclaved meat homogenates, results indicated that the microflora present in Brand B may be the reason for high metabolic activities. Based on the denaturing gradient gel electrophoresis and the Shannon–Wiener diversity index analysis, the “Brand” factor significantly impacted the diversity index (P = 0.012) and Brand B had the highest microflora diversity (Shannon index 1.636 ± 0.011). Based on this study, results showed that antimicrobials cannot completely inhibit the growth of L. monocytogenes in ready‐to‐eat roast beef. Native microflora (both diversity and abundance), together with product formula, pH, antimicrobial concentrations, and storage conditions may all impact the survival and growth of L. monocytogenes.     

Effects of UV‐C on antioxidant capacity,antioxidant enzyme activity and colour of fresh‐cut red cabbage during storage

Ultraviolet C (UV‐C) irradiation is a widely used nonthermal sterilisation method used to ensure the quality and safety of fresh and fresh‐cut produce. However, research focus of UV‐C has recently shifted from safety of sterilisation damage towards improvements in quality. Red cabbage (Brassica oleracea var. capitata f. rubra) is a widely consumed fresh‐cut vegetable. In this study, the effects of UV‐C irradiation on the antioxidant capacity, antioxidant enzyme activity and colour of fresh‐cut red cabbage were evaluated. The UV‐C decreased the L*, a* and b* values, and it turned the colour darker and increasingly blue. UV‐C increased the activity of four antioxidant enzymes and also stimulated the accumulation of flavonoids, glutathione and ascorbic acids. The antioxidant activity of UV‐C treatment was higher than in the control group. These results suggest that an appropriate dose of UV‐C may improve the antioxidant properties of fresh‐cut red cabbage.     

Application of a molecular beacon-real-time PCR technology to detect Salmonella species contaminating fruits and vegetables

An oligonucleotide probe that becomes fluorescent upon hybridization to the target DNA (molecular beacon; MB) was evaluated in a real-time polymerase chain reaction (PCR) assay to detect the presence of Salmonella species. As few as 1-4 colony-forming units (CFU) per PCR reaction could be detected. The capability of the assay to detect Salmonella species from artificially inoculated fresh-cut produce such as cantaloupe, mixed-salad, cilantro, and alfalfa sprouts was demonstrated. A comparison of two commercially available kits utilizing MB-PCR (iQ-Check, Bio-Rad Laboratories) and conventional Association of Official Analytical Chemists (AOAC)-approved PCR (BAX, Dupont Qualicon) was performed on artificially inoculated produce. As few as 4 CFU/25 g of produce were detected after 16 h of enrichment in buffered peptone broth. These assays could be carried out entirely in sealed PCR tubes, enabling a rapid and high-throughput detection of Salmonella species in a large number of food and environmental samples. This is the first report of the application of MB probe being used for real-time detection of Salmonella species in whole and fresh-cut fruits and vegetables.     

Effects of Antimicrobial Coatings and Cryogenic Freezing on Survival and Growth of Listeria innocua on Frozen Ready‐to‐Eat Shrimp during Thawing

Foodborne pathogens such as Listeria monocytogenes could pose a health risk on frozen ready‐to‐eat (RTE) shrimp as the pathogen could grow following thawing. In this study, antimicrobial‐coating treatments alone, or in combination with cryogenic freezing, were evaluated for their ability to inhibit the growth of Listeria innocua, a surrogate for L. monocytogenes, on RTE shrimp. Cooked RTE shrimp were inoculated with L. innocua at 3 population levels and treated with coating solutions consisting of chitosan, allyl isothiocyanate (AIT), or lauric arginate ester (LAE). The treated shrimp were then stored at –18 °C for 6 d before being thawed at 4, 10, or 22 °C for either 24 or 48 h. Results revealed that antimicrobial coatings achieved approximately 5.5 to 1 log CFU/g reduction of L. innocua on RTE shrimp after the treatments, depending on the inoculated population levels. The coating‐treated shrimp samples had significantly (P < 0.05) less L. innocua than controls at each thawing temperature and time. Cryogenic freezing in combination with coating treatments did not achieve synergistic effects against L. innocua. Antimicrobial coatings can help to improve product safety by reducing Listeria on RTE shrimp.     

Control of Listeria monocytogenes Contamination in Ready‐to‐Eat Meat Products

The ubiquitous nature of Listeria monocytogenes and its ability to grow at refrigerated temperature makes L. monocytogenes a significant threat to the safety of ready‐to‐eat (RTE) meat products. The contamination by L. monocytogenes in RTE meat primarily occurs during slicing and packaging after cooking. The effectiveness of post‐package decontamination technology such as in‐package thermal pasteurization, irradiation, and high‐pressure processing are discussed. Formulating meat products with antimicrobial additives is another common approach to control L. monocytogenes in RTE meat. Irradiation is an effective technology to eliminate L. monocytogenes but can influence the quality of RTE meat products significantly. The effect of irradiation or the combination of irradiation and antimicrobials on the survival of L. monocytogenes and the quality of RTE meat is discussed.     

Efficacy of Combined Sous Vide‐Microwave Cooking for Foodborne Pathogen Inactivation in Ready‐to‐Eat Chicory Stems

There is a variety of different food processing methods, which can be used to prepare ready‐to‐eat foods. However, the need to preserve the freshness and nutritional qualities leads to the application of mild technologies which may be insufficient to inactivate microbial pathogens. In this work, fresh chicory stems were packed under a vacuum in films, which were transparent to microwaves. These were then exposed to microwaves for different periods of time. The application of sous vide microwave cooking (SV‐MW, 900 W, 2450 MHz), controlled naturally occurring mesophilic aerobic bacteria, yeasts and molds for up to 30 d when vacuum‐packed vegetables were stored at 4 °C. In addition, the process lethality of the SV‐MW 90 s cooking was experimentally validated. This treatment led to 6.07 ± 0.7 and 4.92 ± 0.65 log cfu/g reduction of Escherichia coli and Listeria monocytogenes inoculated over the chicory stems (100 g), respectively. With an initial load of 9 log cfu/g for both pathogens, less than 10 cfu/g of surviving cells were found after 90 s cooking. This shows that short‐time microwave cooking can be used to effectively pasteurize vacuum‐packed chicory stems, achieving >5 log cfu/g reduction of E. coli and L. monocytogenes.     

Investigation of Listeria monocytogenes contamination pattern in local Chinese food market and the tracing of two clinical isolates by RAPD analysis

This study was undertaken to investigate the contamination pattern of Listeria monocytogenes in local Chinese food markets and to trace two clinical isolates. Random amplification polymorphic DNA (RAPD) was developed to track the source of L. monocytogenes in ready-to-eat (RTE) food products and from clinical origin. Three random primers, PB1, PB4 and HLWL74, were used to subtype all the L. monocytogenes strains isolated from RTE food products, fresh food products, environmental sewages in the markets and two clinical meningitis patients. It was shown that all the 49 isolates could be classified into 5, 4 and 4 types using these three random primers, PB1, PB4, and HLWL74, respectively. Twenty-seven composite profiles were identified by a combination of the three primers. The same composite profiles of L. monocytogenes could be found both in the fresh food products, environmental sewages and RTE food products, suggesting that the L. monocytogenes in the RTE food products may come from those in the fresh food products or sewage in the same market. The composite profiles of the two clinical isolates were the same as those of strains isolated from RTE food products, indicating that the disease might have resulted from the consumption of the RTE food products contaminated with L. monocytogenes. The results show that RAPD could be a powerful tool for the investigation of contamination pattern of L. monocytogenes in Chinese food markets and also for tracking the source of L. monocytogenes in clinical patients.     

A STUDY ON SUITABILITY OF FOUR ENRICHMENT BROTHS FOR PCR-BASED DETECTION OF LISTERIA MONOCYTOGENES FROM RAW MEAT

Four enrichment broths were evaluated for their compatibility with the polymerase chain reaction (PCR) for detection of Listeria monocytogenes from raw meat after single‐step enrichment. Standardized PCR protocols for listeriolysin O (hlyA) gene were used for the species‐specific identification of L. monocytogenes. Four broths, namely, modified University of Vermont broth (MUVM), Listeria enrichment broth (LEB), Fraser broth (FB) and polymyxin, acriflavin, lithium chloride, ceftazidime, aesculin, mannitol, egg yolk broth (PALCAM) , were inoculated with L. monocytogenes. The enriched cultures were subjected for PCR. Similarly, meat samples were artificially spiked with various concentrations of L. monocytogenes, these spiked samples were enriched in the above‐mentioned four broths and subjected to PCR to determine the medium that was most compatible for PCR‐based detection of L. monocytogenes. The aliquots taken during different incubation periods were subjected to three different procedures for the concentration of the target organism for use in PCR. Results revealed that MUVM was better than other broths for the detection of L. monocytogenes by both PCR and cultural method; moreover, it was able to support the growth of as low as 10 cfu/g of meat. Concentration of the target organisms by centrifugation and washing with PCR buffer was the most suitable method for improving PCR performance for detection of L. monocytogenes. Goat (n = 67) and buffalo (n = 45) meat samples from local markets were also screened by both PCR and cultural method to validate the results obtained from the spiking studies. Both results were in agreement in spiking studies as well as screening of market meat samples.     

Reliability of Listeria monocytogenes Identification by Specific PCR Assessed by Phenotypic and Genotypic Techniques

This study evaluates the possibility of using polymerase chain reaction (PCR) for rapid identification of food-borne Listeria monocytogenes as an alternative to API Listeria system and estimates the incidence of API Listeria misidentifications in food-borne Listeria species. A total of 198 strains, 11 L. monocytogenes, 28 other Listeria species, and 159 food isolates were phenotypically and genotypically characterized by API Listeria profiles and randomly amplified polymorphic DNA (RAPD) profiles, respectively. They were also tested for PCR amplification using genus- and species-specific primers. Clustering analysis of phenotypic and genotypic data showed discrepancies in species identification of some isolates by API Listeria profiles. Their identities were confirmed by 16S rDNA sequencing, and thus, it was revealed that 33% of Listeria innocua and 19% of Listeria welshimeri were misidentified as L. monocytogenes by API Listeria profiles. Reliable identification of L. monocytogenes was obtained by LM1–LM2 specific primers which allowed PCR amplification only in reference strains and isolates previously identified as L. monocytogenes by RAPD and 16S rDNA sequence analysis. These results corroborate the suitability of specific PCR as a rapid and accurate test for the identification of L. monocytogenes, avoiding misidentification with other Listeria species commonly found in food products.     

Effect of Salt Reduction on Growth of Listeria monocytogenes in Meat and Poultry Systems

Abstract: Reducing sodium in food could have an effect on food safety. The objective was to determine differences in growth of Listeria monocytogenes in meat and poultry systems with salt substitutes. For phase 1, fresh ground beef, pork, and turkey with NaCl, KCl, CaCl₂, MgCl₂, sea salt, or replacement salt added at 2.0% were inoculated with L. monocytogenes to determine growth/survival during 5 d at 4 °C to simulate a pre‐blend process. L. monocytogenes populations significantly decreased (0.41 log CFU/g) during the storage time in beef, but no differences (P > 0.05) were observed over time in pork or turkey. Salt type did not affect (P > 0.05) L. monocytogenes populations during pre‐blend storage. MgCl₂ and NaCl allowed significant growth of aerobic populations during storage. For phase 2, emulsified beef and pork products were processed with 2% NaCl, KCl, sea salt, or a NaCl/KCl blend and post‐process surface‐inoculated with L. monocytogenes to determine growth/survival at 4 °C for 28 d. Pork products showed significantly greater L. monocytogenes population growth at all sampling times (0, 7, 14, 21, and 28 d) than beef products, but salt type had no effect on L. monocytogenes populations with sampling times pooled for data analysis. Although salt types had no impact on L. monocytogenes populations in preblend and emulsified meat products, pork and turkey preblends and emulsified pork had greater L. monocytogenes populations compared with beef products. These studies demonstrate that sodium may not affect the safety of preblends and emulsified meat and poultry products. Practical Application: odium reduction in food is an important topic because of sodium's unfavorable health effects. This research shows that reducing sodium in pre‐blends and emulsified meat and poultry products would have no effect on Listeria monocytogenes populations, but replacement of NaCl with MgCl₂ may affect growth of aerobic populations.     

Predictive Modeling of the Effect of ε‐Polylysine Hydrochloride on Growth and Thermal Inactivation of Listeria monocytogenes in Fish Balls

This study aimed to evaluate the effect of ε‐polylysine hydrochloride (ε‐PLH) on the growth and thermal inactivation kinetics of Listeria monocytogenes in fish balls. Samples, supplemented with ε‐PLH (0, 150, or 300 ppm, w/w), were inoculated with a three‐strain cocktail of L. monocytogenes and incubated at constant temperatures of 3.4, 8, 12, or 16 °C for growth studies, or heated at 60, 62.5, 65, or 67.5 °C for thermal inactivation tests. The growth curves were fitted to the Huang primary model, and the Huang and Ratkowsky square‐root models (SRM) were used as the secondary models to evaluate the effect of temperature on bacterial growth. The survival during heating was analyzed with a log‐linear model. The results showed that, while the lag time of L. monocytogenes was affected by both ε‐PLH concentration and temperature, the specific growth rate was unaffected by ε‐PLH. Under the same temperature, a 10‐time in increase of the lag time would be expected for every 565 ppm in the increase of ε‐PLH concentration. Using the Ratkowsky SRM, the estimated nominal minimum growth temperature was –2.04 °C, while the minimum growth temperature was 0.29 °C when estimated with the Huang SRM. Validation at 10 °C showed that the Huang primary model, in combination with either the Huang or Ratkowsky SRM, could accurately predict the growth of L. monocytogenes. On the other hand, the thermal resistance of the pathogen was significantly reduced by increase in temperature or ε‐PLH. The thermal z value of L. monocytogenes was 5.78 °C, and the ε‐PLH z value was 1642 ppm. The results of this study showed that the combined application of ε‐PLH and temperature can be used to control L. monocytogenes in fish balls and to improve food safety and reduce risks to public health.     

Microbiological Quality and Safety of Fresh Fruits and Vegetables at Retail Levels in Korea

The aim of this study was to evaluate the microbiological quality and safety of fresh produce at retail level in Korea in order to periodically update information and establish available risks associated with consumption of fresh fruits and vegetables. The samples from different markets located in 3 provinces of South Korea were collected. The protocol in the Korean Food Standards Codex was applied and generic Escherichia coli, coliforms, aerobic mesophilic bacteria (AMB), and yeast and mold (YM) in 360 packaged and unpackaged fresh fruits and vegetables were analyzed. Presence of pathogens was examined using real‐time polymerase chain reaction (q‐PCR) after enrichment of samples. For all, the microbial counts ranged from 1.7 to 10.6 log cfu/g for AMB, 2.2 to 7.9 log cfu/g for coliforms, and 5.5 to 7.9 log cfu/g for YM. Three lettuce samples were contaminated by E. coli with a bacterial load ranging from 2 to 4 log cfu/g. Salmonella spp. were not detected in any fresh produce. Listeria monocytogenes, E. coli O157:H7, and Staphylococcus aureus were found in 1 (0.6%), 3 (0.8%), and 5 (1.4%) fresh produce samples, respectively. Bacillus cereus (50.3%) and Clostridium perfringens (13.3%) had the highest prevalence. These results indicate the need for employing strict control measures and developing preventive strategies to improve the quality and safety of fresh produce in Korea.     

Development and application of a new multiplex real‐time PCR assay for simultaneous identification of Brucella melitensis,Cronobacter sakazakii and Listeria monocytogenes in raw milk and cheese

Brucella melitensis, Cronobacter sakazakii and Listeria monocytogenes are important foodborne pathogens in milk and milk products, which are responsible for a variety of diseases that pose serious hazards to public health and food safety. The objective of this study was to develop a novel multiplex RTi‐PCR for the detection of B. melitensis, C. sakazakii and L. monocytogenes and to characterise the potential risk of these pathogens in raw milk and cheese. The raw milk (n = 25) and cheese samples (n = 20) were analysed by multiplex RTi‐PCR assay and detected for quantification of the three pathogens. In this study, B. melitensis, C. sakazakii and L. monocytogenes were simultaneously identified using BMEII0466, mms operon and hly as target genes, respectively. The multiplex RTi‐PCR assay that was developed showed good sensitivity and selectivity for the pathogenic microorganisms (r² = 0.986–0.997). Multiplex RTi‐PCR results showed that most of the samples were contaminated with the pathogens screened.